| Description | SpeciesGene IDAccessionECSourceLengthMWTag2019-nCoV43740578YP_009725301.13.4.22.69E. coli306aa~33.8kDa-About this proteinName2019-nCoV Main ProteaseSynonyms2019-nCoV Main Protease (Mpro)/ 3C-like Protease (3CLpro), Mpro/3CLpro, 2019-nCoV 3C-like Proteinase, main proteinase, 3C-like proteinase (NSP5)SpeciesGene IDAccessionECSourceLengthMWTag2019-nCoV43740578YP_009725301.13.4.22.69E. coli306aa~33.8kDa-About this proteinName2019-nCoV Main ProteaseSynonyms2019-nCoV Main Protease (Mpro)/ 3C-like Protease (3CLpro), Mpro/3CLpro, 2019-nCoV 3C-like Proteinase, main proteinase, 3C-like proteinase (NSP5), Coronavirus endopeptidase C30, chymotrypsin-like protease, coronavirus 3C-like protease, coronavirus M(pro), coronavirus main proteaseBackground2019-nCoV is a positive-sense single-stranded RNA virus and has high homology with SARS-CoV and MERS-CoV. After infecting host cells, the coronaviral ORF1a/b RNA is first translated with the help of host cells into two polyprotein precursors (PP1a and PP1ab) which are then cleaved by Mpro and papain like proteases to produce multiple nonstructural proteins. These nonstructural proteins are involved in the production of viral subgene RNA and four structural proteins (envelope/E protein, membrane/M protein, spike/S protein and nucleocapsid/N protein), which in turn complete the reproduction and release of the progeny virus. Since the Mpro protease plays a vital role in the life cycle of the virus and there is no homologous protein in the human body, the Mpro main protease is an ideal target for developing antiviral drugs. Meanwhile because Mpro is highly conservative among beta coronaviruses, the Mpro inhibitors selected have a broad-spectrum anti-coronavirus ability and may even be used in curing other animal diseases such as porcine coronavirus.Aladdin's 2019-nCoV Main Protease is purified using the PerfectProtein™ Platform developed by aladdin. It is recombinantly expressed in E. coli without any additional tags or amino acids, ensuring complete identity with the natural Main Protease in 2019-nCoV virus.s2019-nCoV inhibitor screening; enzyme activity or structural studiesPhysical AppearanceliquidBiological Activity≥300U/g or 300µmol/min/gUnit DefinitionOne unit is defined as the amount of enzyme that catalyzes the production of 1µmol of MCA-AVLQ per minute at 37℃ under pH7.0. Concentration~1mg/mlPurity≥ 95% by SDS-PAGE.Formulation500mM Tris, 150mM NaCl, 1mM EDTA, 50% Glycerol, pH7.3Recommended UsageFor inhibitor screening, add 0.2-1µg of this product generally in a reaction volume of 200µl.Amino Acid SequenceSGFRKMAFPS GKVEGCMVQV TCGTTTLNGL WLDDVVYCPR HVICTSEDML NPNYEDLLIR KSNHNFLVQA GNVQLRVIGH SMQNCVLKLK VDTANPKTPK YKFVRIQPGQ TFSVLACYNG SPSGVYQCAM RPNFTIKGSF LNGSCGSVGF NIDYDCVSFC YMHHMELPTG VHAGTDLEGN FYGPFVDRQT AQAAGTDTTI TVNVLAWLYA AVINGDRWFL NRFTTTLNDF NLVAMKYNYE PLTQDHVDIL GPLSAQTGIA VLDMCASLKE LLQNGMNGRT ILGSALLEDE FTPFDVVRQC SGVTFQNotePrecautions：Before opening the tube, centrifuge briefly to collect liquid at the bottom of the tube.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.Instructions for Use：1. For the usage of this product, please refer to the manual of ’s Coronavirus Mpro/3CLpro Activity Fluorometric Assay Kit or other relevant literature, depending on the experimental purposes.2. Please refer to Figure 1 for the performance of this product in cleaving the CoV Main Protease Fluorogenic Substrate, MCA-AVLQSGFR-Lys(Dnp)-Lys-NH2, into a fluorescent product with Ex/Em of 325/393nm.Figure 1. Digestion of MCA-AVLQSGFR-Lys(Dnp)-Lys-NH2 by different amounts of 2019-nCoV Main Protease (Mpro/3CLpro) (, P2320). This figure is for reference only, which may vary due to different experimental conditions... Read More | Inquire | Inquire | Taq MasterMix is a premixed system composed of Taq DNA Polymerase, Mg2+, dNTPs, as well as PCR stabilizers and enhancers, with a concentration of 2 ×. The pre prepared PCR mixture makes the operation simpler and faster, and can minimize human error and contamination to the greatest extent Taq MasterMix is a premixed system composed of Taq DNA Polymerase, Mg2+, dNTPs, as well as PCR stabilizers and enhancers, with a concentration of 2 ×. The pre prepared PCR mixture makes the operation simpler and faster, and can minimize human error and contamination to the greatest extent possible. The original MasterMix formula results in high yield, strong repeatability, and good stability of amplified products. This product has been added with a dye (blue), and can be directly subjected to electrophoresis detection after the reaction is completed. The amplified PCR product has an "A" base attached to its 3 'end, making it suitable for direct use in T/A cloning. Mainly suitable for PCR amplification of DNA, DNA sequencing and other experiments. T665590Component5 mL25 mLStorageT665590A2×Taq MasterMix (Dye)5×1 mL5×5 mL-20℃. Avoid freeze/thaw cycle.T665590BddH₂O5×1 mL5×5 mL-20℃. Avoid freeze/thaw cycle.2×Taq MasterMix contains Taq DNA Polymerase, 3 mM Mg Cl₂ and 400 µM each dNTP. Quality control:After testing, there was no exogenous nuclease activity; PCR method for detecting residual DNA without host; Can effectively amplify single copy genes from multiple genomes.Usage:The following is an example of a PCR reaction system and reaction conditions for amplifying a 1 kb fragment using human genomic DNA as a template. In practical operation, corresponding improvements and optimizations should be made based on the template, primer structure, and target fragment size.1. PCR reaction system Reagent 50 µlReaction system Final concentration 2×Taq MasterMix（Dye） 25 µL 1× Forward Primer，10 µM 2 µL 0.4 µM Reverse Primer，10 µM 2 µl 0.4 µM Template DNA <0.5 µg <0.5 µg/50 µL ddH2O up to 50 µL /Attention: The primer concentration should be between 0.1 and 1.0 as the final concentration µ M serves as a reference for setting the range. In the case of low amplification efficiency, the concentration of primers can be increased; When non-specific reactions occur, the primer concentration can be reduced to optimize the reaction system.2. PCR reaction conditions Step Temperature Time / Pre denaturation 94℃ 2 min / Denaturation 94℃ 30 s 25-35 cycles Anneal 55-65℃ 30 s 25-35 cycles Extend 72℃ 30 s 25-35 cycles Finally extended 72℃ 2 min / Attention:1) In general experiments, if the annealing temperature is 5 ° C lower than the melting temperature Tm of the amplification primer, and the ideal amplification efficiency cannot be achieved, the annealing temperature should be appropriately reduced; When non-specific reactions occur, increase the annealing temperature to optimize the reaction conditions.2) The extension time should be set according to the size of the amplified fragment. The amplification efficiency of Taq DNA Polymerase in this product is 2 kb/min.3) The number of cycles can be set based on the downstream application of the amplification product. If the number of cycles is too small, the amplification amount is insufficient; If there are too many cycles, the probability of mismatches will increase, and non-specific backgrounds will be severe. So, while ensuring product yield, the number of cycles should be minimized as much as possible... Read More | Inquire |